CN102432670A - Silkworm chrysalis protein source dipeptide SS and application thereof - Google Patents
Silkworm chrysalis protein source dipeptide SS and application thereof Download PDFInfo
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- CN102432670A CN102432670A CN2011103861901A CN201110386190A CN102432670A CN 102432670 A CN102432670 A CN 102432670A CN 2011103861901 A CN2011103861901 A CN 2011103861901A CN 201110386190 A CN201110386190 A CN 201110386190A CN 102432670 A CN102432670 A CN 102432670A
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- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
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- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses silkworm chrysalis protein source dipeptide SS. An amino acid sequence of the silkworm chrysalis protein source dipeptide SS is Ser-Ser. The silkworm chrysalis protein source dipeptide SS can be used as an angiotensin converting enzyme (ACE) inhibitory peptide.
Description
Technical field
The invention belongs to biological technical field, particularly an ability combines with Zinc metallopeptidase Zace1, suppresses its active dipeptides Ser-Ser (SS).
Background technology
Zinc metallopeptidase Zace1 (Angiotensin converting enzyme, ACE, EC3.4.15.1; Former name has kininaseIl in the document, dipeptidyl carboxypeptidase I etc.) be a kind of two carboxypeptidases, be to cause a hypertensive key enzyme; It changes into angiotensin II to angiotonin I through hydrolytic action, meanwhile, but ACE passivation bradykinin also; These two kinds of effects all can cause vasoconstriction, thereby cause hypertension.Therefore ACE is considered to cause a hypertensive important factor.Find angiotensin converting enzyme inhibitor (ACEI) after deliberation, can reach hypotensive effect through the activity that suppresses ACE.ACE inhibitor is widely used in diseases such as treatment is cardiovascular, hypertension, heart failure, renal failure.ACE inhibitor is found from snake venom at first, it is found that the ace inhibitory peptide that from foodstuff raw material, extracts subsequently, all can be used as the raw material of preparation ace inhibitory peptide like gelatin, casein, fish, Fructus Fici natural gum, α-zein etc.
Summary of the invention
The technical problem that the present invention will solve provides a kind of pupa albumen source dipeptides and uses thereof.
In order to solve the problems of the technologies described above, the present invention provides a kind of pupa albumen source dipeptides SS, and the aminoacid sequence of this pupa albumen source dipeptides SS is Ser Ser.
The present invention also provides the purposes of above-mentioned pupa albumen source dipeptides SS simultaneously: as ace inhibitory peptide.
Pupa albumen of the present invention source dipeptides; It derives from pupa albumen Bmb000010 sequence, and this dipeptides is that two amino-acid residues in site such as the 59-60,96-97,251-252,267-268,682-683,886-887,1047-1048,1154-1155,1253-1254,1306-1307 in the above-mentioned sequence are formed.
The acquisition approach of dipeptides of the present invention is:
1, based on the screening of the ace inhibitory peptide of Pharmacophore Model:
Pharmacophore is a kind of method for expressing of the abstract characterization of molecules relevant with biological activity.That these abstract characterization of molecules comprise is three-dimensional (hydrophilic and hydrophobic group, charged/ionogen, hydrogen bond donor/acceptor etc.), (minor structure) and the unidimensional (materialization and biological property) of two dimension.The present invention utilizes 23 dipeptides structures of having reported and ACE to suppress active compound DS with certain difference and makes up three-dimensional Pharmacophore Model (table 1).
Table 1, training set peptide storehouse
| Sequence number | Two peptide sequences | ?IC 50(μmol/L) | |
| 1 | VW | ?1.4 | 5.6635 |
| 2 | IY | ?2.1 | 5.2581 |
| 3 | YW | ?10 | 3.6974 |
| 4 | DG | ?12 | 3.5151 |
| 5 | VY | ?10 | 3.6974 |
| 6 | RY | ?10.5 | 3.6486 |
| 7 | AF | ?15.2 | 3.2787 |
| 8 | LY | ?18 | 3.1096 |
| 9 | FY | ?25 | 2.7811 |
| 10 | VK | ?13 | 3.4351 |
| 11 | MF | ?45 | 2.1933 |
| 12 | LW | ?50 | 2.0880 |
| 13 | YN | ?51 | 2.0682 |
| 14 | SF | ?130 | 1.1325 |
| 15 | GY | ?259 | 0.4432 |
| 16 | FP | ?315 | 0.2474 |
| 17 | LF | ?349 | 0.1449 |
| 18 | IR | ?696 | -0.5453 |
| 19 | YP | ?720 | -0.5793 |
| 20 | QK | ?885 | -0.7856 |
| 21 | GK | ?2500 | -1.8240 |
| 22 | GS | ?3800 | -2.2428 |
| 23 | SG | ?8500 | -3.0478 |
The data processing software that the present invention adopted is MOE (Molecular Operating Environment), through the AUTOGPA module of MOE software training set is carried out quantitative structure and imitates analysis, the superimposed result of the molecule of gained training set such as Fig. 1.The Pharmacophore Model that obtains training set from superimposed result is: 2 hydrogen bond receptors, 1 aromatic nucleus center and 1 hydrogen-bond donor (like Fig. 2).Through above-mentioned gained Pharmacophore Model, carry out the activity prediction of verification msg collection, the predictive ability of checking Pharmacophore Model, the result is through the active checking of the ACE of 10 dipeptides structures of having reported, and the result proves the r of model
2=0.8378, explain that Pharmacophore Model has the ideal predictive ability, uses pIC
50Value is represented the size of activity value, pIC
50=6-logIC
50(like table 2 and Fig. 3).
Table 2 checking Ji Taiku
| Sequence number | Two peptide sequences | ?IC 50(μmol/L) | pIC 50 | $ |
| 1 | VW | ?1.4 | 5.6635 | 5.7873 |
| 2 | AW | ?15.4 | 3.2656 | 4.9016 |
| 3 | AA | ?51.4 | 2.0604 | 0.7336 |
| 4 | GP | ?66 | 1.8103 | -0.9967 |
| 5 | AP | ?270 | 0.4016 | 0.8752 |
| 6 | FP | ?315 | 0.2474 | 0.5412 |
| 7 | AG | ?2500 | -1.8240 | -0.5164 |
| 8 | GK | ?5400 | -2.5942 | -2.9616 |
| 9 | AH | ?9000 | -3.1050 | -4.9316 |
| 10 | GD | ?9200 | -3.1270 | -3.3091 |
Annotate: $pIC
50What represent is active predictor, with pIC
50The size of measured value is made comparisons.
2, the activity prediction of specific dipeptides in the virtual peptide storehouse:
With the proteic sequence of Bmb000010 input PeptideCutter (
Http:// web.expasy.org/peptide_cutter/) simulation enzymolysis software, all proteolytic enzyme that utilize software to provide carry out complex enzyme hydrolysis, from a series of different lengths peptide of enzymolysis gained section, select dipeptides structure wherein, and SS is one of them.
Through Pharmacophore Model dipeptides SS (according to the invention) is carried out the activity prediction, gained prediction activity is $pIC
50=3.935, corresponding IC
50=7.88 μ mol/L indicate that this dipeptides structure possibly have good ACE and suppress active.
3, ACE suppresses active detection method.
ACE is at 37 ℃, and the pH value is that the stand-in Hippuryl-L-Histidyl-L-Leucine (HHL) of catalytically decomposed angiotensin I produces urobenzoic acid (HA) under 8.3 the condition, and this material has charateristic avsorption band at ultraviolet 225nm place; ACE is suppressed the catalyticing decomposition action of HHL when adding ACE inhibitor, and the growing amount of urobenzoic acid reduces, and through the HPLC method, the variation of the amount of the urobenzoic acid that generates can be calculated and suppressed active size before and after the mensuration adding suppressor factor.
Reaction system is: the ACE, the 50 μ L ace inhibitory peptides (being the SS dipeptides) that add 20 μ L 0.1U/mL are successively respectively bathed 5min 37 ℃ of temperature; The HHL substrate that adds 10 μ L 5mM then starts the catalyzed reaction of ACE; The HCl termination reaction that behind 37 ℃ of shaking bath 30min, adds 250 μ L 1.0moL/L, system solution are crossed the content that carries out RP-HPLC check and analysis urobenzoic acid (HA) behind the 0.45 μ m filter membrane.Above-mentioned similarity condition, (NaCl that contains 0.3moL/L pH=8.3) replaces ACE inhibitor as blank reaction system in the borate buffer with 50 μ L 0.1moL/L.
Annotate: above-mentioned ACE, HHL substrate all are that (NaCl that contains 0.3moL/L pH=8.3) is solvent for borate buffer with 0.1moL/L.
Ace inhibitory peptide (SS dipeptides), be dissolved in different concns in the borate buffer of 0.1moL/L (NaCl that contains 0.3moL/L, pH=8.3) in and get.
RP-HPLC detects: solvent I is that the triethylamine (TTA) of 0.05% (V/V) trifluoroacetic acid (TFA) and 0.05% (v/v) is dissolved in the deionized water, and solvent II is 100% chromatographically pure second eyeball.The ratio of solvent I and solvent II is 70%: 30% (volume ratio), and flow velocity is 0.5mL/min, and the detection wavelength is 225nm, and detecting column temperature is 30 ℃.
It is active in computes that ACE suppresses:
I％＝(A-B)/A×100％
A: the peak area of the urobenzoic acid when not adding the small peptide suppressor factor;
B: the peak area of the urobenzoic acid when adding the small peptide suppressor factor;
The ACE:1U unit definition is that under the standard detection condition, at 37 ℃, (Hippuryl-L-Histidyl-L-Leucine HHL), produces the amount of 1 μ M ACE that urobenzoic acid consumes to catalytic substrate in the 1min time.That is, be the activity unit of ACE.
Advantage of the present invention and positively effect:
1) dipeptides of the present invention has clear and definite target molecules.
Utilization obtains the potential active ace inhibitory peptide that has based on the virtual screening method of pharmacophore, can quicken the invention speed of new bioactive peptide, for hypertension therapeutic provides more active stronger medicine or medicine guide things.
2) utilize the bioactive peptide of area of computer aided virtual screening, can verify its biological activity through chemosynthesis.
This two peptide molecule can suppress the activity of Zinc metallopeptidase Zace1.
According to the described aminoacid sequence of SEQ ID NO:1 of the present invention, can entrust the biochemical (Shanghai) Co., Ltd. of gill synthetic, thereby obtain ace inhibitory peptide of the present invention (or abbreviating dipeptides SS as).
The usage and the consumption of ace inhibitory peptide of the present invention (or abbreviating dipeptides SS as) are following:
Dipeptides of the present invention is an oral type, and consumption is an oral 0.8g (adult), every day 2～3 times.
Description of drawings
Do further explain below in conjunction with the accompanying drawing specific embodiments of the invention.
Fig. 1 is the superimposed figure as a result of the molecule of 23 dipeptides structures in the training set.
Fig. 2 is based on the Pharmacophore Model figure that training set produces; Comprise 1 hydrogen bond receptor, 1 aromatic nucleus center, 1 hydrogen-bond donor.
Fig. 3 is the correlation analysis figure of prediction activity and substantial activity; GPA predictor and 6-logIC
50The correlation coefficient r of actual value=0.9153, r
2=0.8378
Fig. 4 is that dipeptides SS suppresses active color atlas at the ACE of 1.0mg/mL concentration;
Among Fig. 4:
A is blank, and the ACE that does not add dipeptides SS suppresses active color atlas;
B suppresses active color atlas for the ACE of the dipeptides SS of interpolation 1.0mg/mL concentration.
Fig. 5 is that dipeptides SS suppresses active color atlas at the ACE of 70.3mg/mL concentration;
Among Fig. 5:
A is blank, and the ACE that does not add dipeptides SS suppresses active color atlas;
B suppresses active color atlas for the ACE of the dipeptides SS of interpolation 70.3mg/mL concentration.
Embodiment
Chromatographic condition: solvent I is that the triethylamine (TTA) of 0.05% trifluoroacetic acid (TFA) and 0.05% is dissolved in (being to contain the trifluoroacetic acid of 0.5mL and the triethylamine of 0.5mL among every liter of solvent I) in the deionized water, and solvent II is 100% chromatographically pure second eyeball.The ratio of solvent I and solvent II is 70%: 30% (volume ratio), and ultimate3000 wears the peace liquid chromatograph, and chromatographic column is waters Symmetry C
185 μ m, 4.6 * 250mm, flow velocity is 0.5mL/min, sample size 10 μ L, the detection wavelength is 225nm, detecting column temperature is 30 ℃.
Active (detection method is the same), the color atlas (Fig. 4) as a result of detecting.This moment, SS concentration was 1.0mg/mL.
The result:
A: the peak area of the urobenzoic acid when not adding the small peptide suppressor factor is 10.422mAU,
B: the peak area of the urobenzoic acid when adding the small peptide suppressor factor is 2.652mAU.
So it is 74.55% that the ACE of dipeptides SS when 1.0mg/mL suppresses activity.
Embodiment 2, the dipeptides SS ACE under the concentration of 70.3mg/mL suppresses active:
Chromatographic condition: solvent I is that the triethylamine (TTA) of 0.05% trifluoroacetic acid (TFA) and 0.05% is dissolved in the deionized water; Solvent II is 100% chromatographically pure second eyeball.The ratio of solvent I and solvent II is 70%: 30%, and ultimate3000 wears the peace liquid chromatograph, and chromatographic column is waters Symmetry C
185 μ m, 4.6 * 250mm, flow velocity is 0.5mL/min, sample size 10 μ L, the detection wavelength is 225nm, detecting column temperature is 30 ℃.
Detection method: will obtain this dipeptides structure through chemical synthesis, and carry out activity and detect (detection method is the same), color atlas (Fig. 5) as a result.This moment, SS concentration was 70.3mg/mL.
The result:
A: the peak area of the urobenzoic acid when not adding the small peptide suppressor factor is 10.422mAU,
B: the peak area of the urobenzoic acid when adding the small peptide suppressor factor is 0.650mAU.
So it is 93.76% that the ACE of dipeptides SS when 70.3mg/mL suppresses activity.
Through inhibition concentration and the activity data among embodiment 1 and the embodiment 2,, draw the IC of SS through the LOGIT method in the calculating of medicine inhibition concentration
50Value is 0.104mg/mL, both 541 μ mol/L.Gained actual verification activity value is an ideal, and this dipeptides structure is not appeared in the newspapers, and belongs to new ACE and suppresses bioactive peptide.
Dipeptides SS of the present invention, it is all strong that the ACE of these active dipeptides such as specific activity IR, YP, QK, GK, GS, SG suppresses activity.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention and according to this sequence structure of carrying out derive, all should think protection scope of the present invention.
Claims (2)
1. pupa albumen source dipeptides SS, it is characterized in that: the aminoacid sequence of this pupa albumen source dipeptides SS is Ser-Ser.
2. the purposes of pupa albumen according to claim 1 source dipeptides SS is characterized in that: as ace inhibitory peptide.
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| CN 201110386190 CN102432670B (en) | 2011-11-29 | 2011-11-29 | Silkworm chrysalis protein source dipeptide SS and application thereof |
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| CN 201110386190 CN102432670B (en) | 2011-11-29 | 2011-11-29 | Silkworm chrysalis protein source dipeptide SS and application thereof |
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| CN102432670B CN102432670B (en) | 2013-07-10 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739665A (en) * | 2013-12-30 | 2014-04-23 | 浙江树人大学 | Dipeptide TE with dual functions of lowering blood pressure and lowering blood fat and application thereof |
| WO2016023150A1 (en) * | 2014-08-11 | 2016-02-18 | 广州世优生物科技有限公司 | Application of dipeptide as ace enzyme activity inhibitor |
| CN108701171A (en) * | 2015-10-22 | 2018-10-23 | 马古苏托科技大学 | In pharmacophore, the Compounds and methods for by inhibiting that there is application in CYP17A1 and CYP19A1 treating cancers |
| WO2019006951A1 (en) * | 2017-07-07 | 2019-01-10 | 广州世优生物科技有限公司 | Application of nonpolar dipeptide for preparing antihypertensive drug or health care product |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000191690A (en) * | 1998-05-29 | 2000-07-11 | Ichiban Shokuhin Kk | Angiotensin converting enzyme inhibitory peptide |
-
2011
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000191690A (en) * | 1998-05-29 | 2000-07-11 | Ichiban Shokuhin Kk | Angiotensin converting enzyme inhibitory peptide |
Non-Patent Citations (3)
| Title |
|---|
| WANG Z: "Angiotensin-1-converting enzyme inhibitory peptides:Chemical feature based pharmacophore generation", 《EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY》 * |
| 周宁: "液相色谱-质谱联用技术分析三氯氧磷辅助下丝氨酸和组氨酸的成肽产物", 《厦门大学学报(自然科学版)》 * |
| 王伟: "一种源于蚕蛹蛋白组分的新活性肽分离鉴定及其与ACE作用模式研究", 《中国食品科学技术学会第七届年会论文摘要集》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739665A (en) * | 2013-12-30 | 2014-04-23 | 浙江树人大学 | Dipeptide TE with dual functions of lowering blood pressure and lowering blood fat and application thereof |
| WO2016023150A1 (en) * | 2014-08-11 | 2016-02-18 | 广州世优生物科技有限公司 | Application of dipeptide as ace enzyme activity inhibitor |
| CN106573035A (en) * | 2014-08-11 | 2017-04-19 | 广州世优生物科技有限公司 | Application of dipeptide as ace enzyme activity inhibitor |
| US10441624B2 (en) | 2014-08-11 | 2019-10-15 | Withyou Biotechnology Co., Ltd. | Application of dipeptide as ace enzyme activity inhibitor |
| CN106573035B (en) * | 2014-08-11 | 2020-08-28 | 广州世优生物科技有限公司 | Use of dipeptides as ACE enzyme activity inhibitors |
| CN108701171A (en) * | 2015-10-22 | 2018-10-23 | 马古苏托科技大学 | In pharmacophore, the Compounds and methods for by inhibiting that there is application in CYP17A1 and CYP19A1 treating cancers |
| CN108701171B (en) * | 2015-10-22 | 2022-06-10 | 马古苏托科技大学 | Pharmacophores, compounds and methods having application in the treatment of cancer by inhibition of CYP17a1 and CYP19a1 |
| WO2019006951A1 (en) * | 2017-07-07 | 2019-01-10 | 广州世优生物科技有限公司 | Application of nonpolar dipeptide for preparing antihypertensive drug or health care product |
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| Publication number | Publication date |
|---|---|
| CN102432670B (en) | 2013-07-10 |
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